Objective To study the inhibitory effects of adenovirus-mediated fusion gene system driven by KDR promoter(AdKDR-CDglyTK) on angiogenesis and growth of pancreatic cancer.Methods The nude mice model was reproduced bearing pancreatic cancer cell lines Capan-2.Twenty nude mice were randomly and equally divided into four groups: AdKDR-CDglyTK and 5-FC/GCV were injected to the animals of group Ⅰ;group Ⅱ received 5-FC/GCV injection,group Ⅲ received AdKDR-CDglyTK injection,and group Ⅳ as control,received neither AdKDR-CDglyTK nor 5-FC/GCV.AdKDR-CDglyTK was injected directly into the tumor,while 5-FC/GCV was given by intraperitoneal injection.The treatment efficiency in each group was observed and the tumor microvessel density(MVD) was analyzed.RT-PCR was employed to examine the expression of CDglyTK in tumors.Results Tumor growth was dramatically inhibited in group Ⅰ,while no significant difference was found in group Ⅱ,group Ⅲ and group Ⅳ.The MVD in the four groups were 2.08?0.79,10.01?0.77,9.91?0.63 and 10.39?1.35,respectively(F=93.29,P=0.00).The MVD decreased significantly in group Ⅰ compared to the other three groups(P0.05).RT-PCR showed that a 2.4kB fragment had been amplified in the tumor tissues of groupⅠand group Ⅲ,but not in groupⅡand group Ⅳ.Conclusion AdKDR-CDglyTK with 5-FC and GCV can significantly inhibit the angiogenesis and growth of implanted pancreatic cancer in nude mice.

OBJECTIVE:To provide reference for rational use of Shexiang tongxin dripping pills in clinic. METHODS:In retrospective study,the utilization of Shexiang tongxin dripping pills in outpatients and inpatients of our hospital during Jan. 2014-May 2015 was analyzed statistically in terms of gender,age,department,irrational drug use,ADR and DUI,etc. RE-SULTS:There were 719 outpatient prescriptions and 281 inpatient medical orders. The female was more than male with ratio of 1.42:1. Most patients aged 80-89 year-old(35.4%). Shexiang tongxin dripping pills were mostly used in cardiovascular medicine de-partment(760 pieces/copies,76.0%). There were 72 prescriptions/medical orders that did not conform to the indications(7.2%), and 7 prescriptions/medical orders for overdose use(0.7%). No obvious ADR was found. DUI of Shexiang tongxin dripping pills was 1.007. CONCLUSIONS:Although the clinical use of Shexiang tongxin dripping pills in our hospital is basically rational,there still are some nonstandard uses,which should be taken seriously. In order to reduce ADR,the clinical use of Shexiang tongxin dripping pills should be strictly in accordance with the drug instructions and should emphasize the syndrome differentiation based on treatment.

BACKGROUND:Liushen Pil is a traditional Chinese herbal formula, has the effects of heat-clearing and detoxicating, eliminating stagnation, detumescence and al eviating pain. Modern pharmacology verifies that Liushen Pil has anti-inflammatory, analgesic, cardiac, anti-viral, anti-tumor effects, and has been extensively used in the treatment of various infectious diseases and malignant cancer. OBJECTIVE:To investigate the inhibitory effects of Liushen Pil on esophageal cancer xenografts, and effects on microvessel density and vascular endothelial growth factor expression. METHODS:After reproducing nude mouse models of human esophageal cancer, 48 nude mice were randomly divided into high-dose Liushen Pil group, moderate-dose Liushen Pil group, low-dose Liushen Pil group, cisplatin group, model group and blank group. According to medication regimen, drugs were given. The growth of transplanted tumor of nude mice was dynamical y observed in each group. The nude mice were sacrificed after 20 days of treatment. Neoplasm weight was taken and the tumor-suppressing rate was calculated. Immunohistochemistry was used to detect microvessel density and the expression of vascular endothelial growth factor. RESULTS AND CONCLUSION:The weight of transplanted tumor was significantly lower in the high-dose Liushen Pil group, moderate-dose Liushen Pil group, low-dose Liushen Pil group, and cisplatin group than in the model group (P<0.05). Microvessel density and the expression of vascular endothelial growth factor were obviously lower in the each Liushen Pil group than in the model group, but not as apparent as that in the cisplatin group. Results suggested that Liushen Pil can inhibit the growth of the esophageal cancer xenografts. Liushen Pil can down-regulate the expression of vascular endothelial growth factor and reduce microvessel density, which is one of the tumor-inhibiting mechanism of Liushen Pil .

Objective To investigate the feasibility, safety and effectiveness of stent-assisted coil embolization for the treatment of intracranial wide-necked aneurysms. Methods The clinical and imaging data of the patients with intracranial wide-necked aneurysm treated with stent-assisted coil embolization were analyzed retrospectively. Results A total of 200 patients with 205 aneurysms were enrolled. The mortality was 1. 5% and the disability rate was 1. 0% at discharge. One hundred seventy-seven patients were followed up for 16-51 months. The modified Rankin Scale scores: 0 in 174 cases, 2 in 2 cases, 4 in 1 case. Eleven patients (5. 5% ) had perioperative complications, including intraoperative bleeding in 3 cases, postoperative bleeding in 3 cases, postoperative cerebral infarction in 2 cases, coil protrusion in 2 cases, and postoperative epileptic seizure in 1 case. Univariate analysis showed that there were significant differences in the proportions of male patients (9. 1% vs. 5. 3% ; χ2 = 4. 42, P = 0. 026), hypertension (54. 5% vs. 23. 3% ; χ2 = 5. 42, P = 0. 03) and stent prior to coil implantation (54. 5% vs. 85. 1% ; χ2 = 3. 54, P =0. 021) between the complication group and the noncomplication group. Multivariate logistic regression analysis showed that the pre-stenting was an independent protective factor for surgery-related complications (odds ratio [OR] 0. 208, 95% confidence interval [CI] 0. 055-0. 791; P = 0. 021), and hypertension was an independent risk factor for surgery-related complications (OR 4. 380, 95% CI 1. 170-16. 399; P = 0. 028). The imaging follow-up of 167 aneurysms was obtained, including 26 recurrent aneurysms (15. 6% ). Univariate analysis showed that there was significant difference in the aneurysm site (anterior circulation aneurysms: 73. 1% vs. 89. 1% ; posterior circulation aneurysms: 26. 9% vs. 10. 6% ; χ2 = 5. 09, P = 0. 033) and size (giant aneurysms: 7. 7% vs. 0. 7% ; large artery aneurysm: 65. 4% vs. 29. 1% ; small aneurysms:26. 9% vs. 70. 2% ; χ2 = 20. 77, P < 0. 001) between the recurrence group and the nonrecurrence group. Multivariate logistic regression analysis showed that large aneurysms (OR 6. 057, 95% CI 2. 296-5. 983; P <0. 001), giant aneurysms (OR 25. 260, 95% CI 1. 903- 335. 267; P = 0. 014 ), and posterior circulation aneurysms ( OR 3. 184, 95% CI 1. 028- 9. 857; P = 0. 045 ) were the independent risk factors of postoperative recurrence. Conclusions Stent-assisted coil embolization is one of the effective methods for the treatment of complex wide-neck aneurysms. Hypertension and coils prior to stenting are the independent risk factors for perioperative complications, and larger aneurysm size and aneurysms in the posterior circulation are the independent risk factors for postoperative recurrence.

BACKGROUND: Recent studies indicate that bone marrow messenchymal stem cells (MSCs) can repairinjured myocardium and improve cardiac functions. However, there are still no effective non-invasive methods to detect ventricular wall motion, survival status of transplanted cells, and microcirculation improvement after MSCsimplantation. OBJECTIVE: To detect the effects of MSCs implantation on the cardiac function of ischemic myoeardium and investigate changes of tissue acoustics characteristics by using integrated backscatter (IBS) and Doppler tissue imaging (DT1). DESIGN, TIME AND SETTING: Completely randomized grouping design, and randomized controlled study, which was carried out in the Department of Cardiology, Renmin Hospital of Wuhan University from December 2004 to June 2005 (important laboratory of 211 government). MATERIALS: Forty-eight male rabbits with irrespective of gender were used for study. Anti-troponin T (TnT) antibody was provided by Maxim Company, USA; anti-bromodeoxyuridine (BrdU) antibody by Sigma Company, USA; Sonos 5500 echocardiography by HP Company, USA.METHODS: Forty-eight rabbits were randomly divided into model group and implantation group. Model of myocardium infarction was created in rabbits by left anterior descending coronary artery (LAD) ligation. Three days after modeling, autologous bone marrow MSCs were injected into crossed myocardium between infarction and non-infarction regions based on 5 injected points.MAIN OUTCOME MEASURES: Indicators including ventricular size, ventricular wall thickness, and ejection fraction (EF) in the left ventricle (LV) were measured with routine echocardiography one day before myocardial infarction, two weeks after myocardial infarction, and four weeks after implantation. IBS and DTI might detect the anterior backscatter parameters (AII, or IB) and ventricular wall motion parameters (PPI, or CVIB) to calculate the average backscatter scores and backscatter cycle changes. DTI was used to measure peak systolic flow velocity (VS), peak early diastolic flow velocity (VE), and peak late diastolic flow velocity (VA) of the middle and basal segments of the anterior wall of left ventricle. Finally, animals were sacrificed by anesthesia, and myocardial infarction region was stained with 5-Brdu immunohistochemical staining, and anti-TnT staining. RESULTS: Twelve rabbits were excluded because of death after operation.① Echocardiography: Imaging of 36 rabbits were clear in 1 day before myocardial infarction, 2 weeks after myocardial infarction, and 4 weeks after implantation. The sizes and systolic function of LV in 36 rabbits were normal before operation. Two weeks after operation, the sizes of LV became larger, the anterior wall of LV became thinner, and systolic range became weaker even disappeared. EF, CVIB, Vs, and Ve were lower (P < 0.05) and All was higher in both groups than pre-operation (P < 0.05). Four weeks after implantation, compared with control group, left ventricular diastolic diameter (LVDd) was smaller and systolic range became larger, All descended. EF, Vs, Ve and CVIB increased (P < 0.05).② Myocardial immunohistochemical detection: Numerous of Brdu-labeled positive cells were scattered, and anti-TnT was positive, which clearly showed horizontal stria of myocardium. However, normal myocardium was negative. CONCLUSION: Bone marrow MSCs after implantation can survive in ischemic myocardium and improve cardiac function. IBS and DTI can real-timely and exactly detect functional changes of infarction myocardium after bone marrow MSCs implantation.

A high-speed counter-current chromatography (HSCCC) method was successfully developed for the preparative separation and purification of deoxyschizandrin from Schisandrae Sphenantherae Fructus in one step. The purity of deoxyschizandrin was 98.5%, and the structure was identified by MS, UV and NMR. This method was simple, fast, convenient and appropriate to prepare pure compound as reference substances for related research on Schisandrae Sphenantherae Fructus.

Objective To investigate the curative effect of the adenovirus-mediated fusion gene system driven by KDR promoter (AdKDR-CDglyTK) on a model of pancreatic cancer. Methods By using transplantation of the cultivated cells, human pancreatic cell line Capan-2 was injected subcutaneously on the back of nude mice to establish the animal model of the pancreatic cancer. Twenty nude mice were divided randomly and equally into four groups. The mice in group Ⅰ were injected with AdKDR-CDglyTK and 5-FC/GCV, those in group Ⅱ were injected with 5-FC/GCV, those in group Ⅲwere injected with AdKDR-CDglyTK and those in group Ⅳ received no any injection. AdKDR-CDglyTK was injected directly into the tumor and 5-FC/GCV was given by intraperitoneal injection. The observing parameters included common status, tumor bulk, tumor weight, inhibition rate of tumor growth, pathology, immunohistochemistry and treatment effect in each group. Electron microscopy was performed to observe the pathological changes of cells. The apoptotic cells in tumor were detected using the TUNEL assay. The expression of CDglyTK in tumors from each group was examined by RT-PCR. Results Tumor growth was dramatically inhibited in group Ⅰ. Tumor growth has no significant difference among groupⅡ , group Ⅲ and group Ⅳ. The apoptotic rate (34.20±4.60)% was significantly increased in group Ⅰ (F= 243. 22, P= 0. 00) and it had no significant difference among groupⅡ , group Ⅲ and group Ⅳ (P＞0.05). Conclusion AdKDR-CDglyTK with 5-FC/GCV can obviously inhibit the growth of human KDR-expressing pancreatic cell line Capan-2 and induce the cell apoptosis in vivo. The probable molecular mechanism lies in the facts that the system can cause a decline in the level of Bcl-2.

Objective To investigate the curative effect of an adenovirus-mediated fusion gene system driven by VEGF promoter (AdVEGF-CDglyTK) on a nude mouse model of colorectal cancer and analyze the mechanism underlying its therapeutic effect.Methods The animal model of the colorectal cancer was established by using transplantation of the cultivated cells,human colorectal cell line LoVo,via subcutaneous injection on the back of nude mice.Twenty nude mice were equally divided into four groups:group Ⅰ received injection of AdVEGF-CDgiyTK plus 5-flurocytosine/ganciclovir(5-FC/GCV);group Ⅱwere given 5-FC/GCV;group Ⅲ were with AdVEGF-CDglyTK;group Ⅳ were used as control.Results CDglyTK was expressed exclusively in the tumor tissues from the group Ⅰ and Ⅲ by RT-PCR.The phenotype and pathological analysis showed that tumor growth was dramatically inhibited in group Ⅰwhen compared with other three groups,while no significant difference was found between group Ⅱ,group Ⅲ and group Ⅳ.The TUNEL assay demonstrated that the apoptosis rate of 38.65% ± 4.20 significantly increased in group Ⅰ when compared with other three groups (F = 397.530,P =0.000).The tumor microvessel density of 3.08±0.79 decreased significantly in group Ⅰ (F = 34.081,P = 0.000) when compared with other three groups.Conclusion The results suggested that AdVEGF-CDglyTK with 5-FC/GCV can inhibit the tumor growth of colorectal cancer significantly in vivo by a mechanism of systeminduced apoptosis and the efficient suppression of angiogenesis.

A reversed phase high performance liquid chromatography （HPLC） method was established for the simultaneous determination of 12, 13-dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture. A Grace Apollo Cl8 column （250 mm × 4.6 mm, 5 μm） was used as the stationary phase and the mobile phase was composed of acetonitrile and aqueous phosphoric acid （0.2%, v/v）. Gradient elution was carried out at the flow rate of 1.0 mL/min and the column temperature was 30 ℃. An ultraviolet （UV） detector was used with a selected wavelength of 240 nm. Calibration curves were linear within the concentration range of 4.6-45.75 μg/mL for 12, 13-dihydroxyeuparin （r〉0.9999） and 106.9-1068.9μg/mL for glycyrrhizic acid （r〉0.9999）, respectively. Recoveries were 102.18% for 12, 13-dihydroxyeuparin and 101.17% for glycyrrhizic acid. The method developed could be applied to the simultaneous determination of 12, 13- dihydroxyeuparin and glycyrrhizic acid in Yanyanfang mixture.